Primary cultured cells were rinsed with PBS three times for five minutes each, fixed in 4% paraformaldehyde/4% sucrose in PBS for 15 minutes, and permeabilized in PBS with 0.3% triton X-100 (PBS-T) for 10 minutes. Non-specific binding of antibodies was blocked by incubating cells with 1% bovine serum albumin (BSA, Sigma-Aldrich) and 5% normal goat serum (Vector Laboratories Inc. Burlingame, CA, USA) in PBS-T for one hour. Cells were stained with primary antibodies overnight at 4°C. After washing three times in PBS five minutes each, cells were incubated with appropriate secondary antibodies (Alexa Fluor® 488 goat anti-rabbit IgG, 1:500, Invitrogen; Alexa Fluor® 555 goat anti-mouse IgG, 1:500, Invitrogen; Alexa Fluor® 488 goat anti-rabbit IgG, 1:500, Invitrogen; Alexa Fluor® 555 goat anti-chicken IgG, 1:1000, Invitrogen) for one hour in the dark. Coverslips were then mounted with DAPI containing media (Vector Laboratories).
Alexa fluor 555 goat anti chicken igg
Manufactured by Thermo Fisher Scientific
Alexa Fluor® 555 goat anti-chicken IgG is a secondary antibody conjugated with the Alexa Fluor® 555 fluorescent dye. It is designed to detect and label chicken immunoglobulin G (IgG) in various immunoassay and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Dapi containing media, by Vector Laboratories (1 mentions)
Normal goat serum (ngs), by Vector Laboratories (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 555 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Chicken anti egfp, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
2 protocols using alexa fluor 555 goat anti chicken igg
1
Immunofluorescence Staining of Cultured Cells
2
Visualizing EGFP Expression in Transfected Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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EGFP
expression from primary neuronal and retinal transfected cells
was examined 72 h after their exposure to nanodiaplexes to qualitatively
assess the transfection efficiency by immunocytochemistry.36 (link) Briefly, cover slips were incubated overnight
with chicken anti-EGFP (Invitrogen, 1:300). Cells were incubated for
1 h with secondary antibody Alexa Fluor555 goat anti-chicken IgG (Invitrogen,
1:100) which was pseudocolored in green to visualize EGFP expression.
Nuclei were stained with Hoechst 33342 (Sigma-Aldrich, Spain). Confocal
images were obtained using a laser-confocal microscope (Leica TCS
SPE Microsystems GmbH, Germany).
expression from primary neuronal and retinal transfected cells
was examined 72 h after their exposure to nanodiaplexes to qualitatively
assess the transfection efficiency by immunocytochemistry.36 (link) Briefly, cover slips were incubated overnight
with chicken anti-EGFP (Invitrogen, 1:300). Cells were incubated for
1 h with secondary antibody Alexa Fluor555 goat anti-chicken IgG (Invitrogen,
1:100) which was pseudocolored in green to visualize EGFP expression.
Nuclei were stained with Hoechst 33342 (Sigma-Aldrich, Spain). Confocal
images were obtained using a laser-confocal microscope (Leica TCS
SPE Microsystems GmbH, Germany).
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